Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water.

Simple item page
dc.contributor.authorTantawiwat, Suwaleeen_US
dc.contributor.authorTansuphasiri, Unchaleeen_US
dc.contributor.authorWongwit, Waranyaen_US
dc.contributor.authorWongchotigul, Vareeen_US
dc.contributor.authorKitayaporn, Dwipen_US
dc.date.accessioned2009-05-27T16:32:06Z
dc.date.available2009-05-27T16:32:06Z
dc.date.issued2005-01-24en_US
dc.descriptionThe Southeast Asian Journal of Tropical Medicine and Public Health.en_US
dc.description.abstractMultiplex PCR amplification of lacZ, uidA and plc genes was developed for the simultaneous detection of total coliform bacteria for Escherichia coli and Clostridium perfringens, in drinking water. Detection by agarose gel electrophoresis yielded a band of 876 bp for the lacZ gene of all coliform bacteria; a band of 147 bp for the uidA gene and a band of 876 bp for the lacZ gene of all strains of E. coli; a band of 280 bp for the p/c gene for all strains of C. perfringens; and a negative result for all three genes when tested with other bacteria. The detection limit was 100 pg for E. coli and C. perfringens, and 1 ng for coliform bacteria when measured with purified DNA. This assay was applied to the detection of these bacteria in spiked water samples. Spiked water samples with 0-1,000 CFU/ml of coliform bacteria and/or E. coli and/or C. perfringens were detected by this multiplex PCR after a pre-enrichment step to increase the sensitivity and to ensure that the detection was based on the presence of cultivable bacteria. The result of bacterial detection from the multiplex PCR was comparable with that of a standard plate count on selective medium (p=0.62). When using standard plate counts as a gold standard, the sensitivity for this test was 99.1% (95% CI 95.33, 99.98) and the specificity was 90.9 % (95% CI 75.67, 98.08). Multiplex PCR amplification with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.en_US
dc.description.affiliationDepartment of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. en7tox@hotmail.comen_US
dc.identifier.citationTantawiwat S, Tansuphasiri U, Wongwit W, Wongchotigul V, Kitayaporn D. Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water. The Southeast Asian Journal of Tropical Medicine and Public Health. 2005 Jan; 36(1): 162-9en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/35430
dc.language.isoengen_US
dc.source.urihttps://www.tm.mahidol.ac.th/seameo/2005_36_1/26-3430.pdfen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshClostridium perfringens --geneticsen_US
dc.subject.meshDNA Primersen_US
dc.subject.meshEnterobacteriaceae --geneticsen_US
dc.subject.meshEnterotoxinsen_US
dc.subject.meshEscherichia coli --geneticsen_US
dc.subject.meshGene Amplificationen_US
dc.subject.meshGenes, Bacterialen_US
dc.subject.meshHumansen_US
dc.subject.meshPolymerase Chain Reaction --methodsen_US
dc.subject.meshSensitivity and Specificityen_US
dc.subject.meshWater Microbiologyen_US
dc.subject.meshWater Supplyen_US
dc.titleDevelopment of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water.en_US
dc.typeIn Vitroen_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
162.pdf
Size:
133.61 KB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.79 KB
Format:
Plain Text
Description:
Download
Collections
Southeast Asian Journal of Tropical Medicine and Public Health